Calcium-sensitive adenylyl cyclases and cAMP compartmentation. (360G-Wellcome-082413_Z_07_Z)

We wish to continue the study of Ca2+-sensitive ACs, along with the cAMP compartmentalization that is an inherent component in their regulation. Since we are beginning to suspect that ACs are no longer just centrally important generators of a critical second messenger but are also dynamic scaffolding platforms around which regulatory assemblies may be gathered, we need to know their partners in functioning signalling complexes. To understand the functions, interactions and regulation in the cell of Ca2+-sensitive ACs we want to extend what we know of the molecular requirements of Ca2+-stimulation and inhibition, by a combination of structure-function, structural and kinetic analyses; we want to know which proteins the ACs associate with, to identify the cellular features that confer Ca2+-sensitivity on ACs and to identify other components of AC microdomains; finally we wish to study cAMP dynamics in these microdomains in real-time in live cells, using rapid cAMP sensors and in situ str ategies, such as RNAi, to unambiguously assess the contribution of individual components in controlling cAMP. We feel that these three interdigitated experimental approaches provide the necessary powerful strategy to bring a true grasp of the complexity, flexibility and power that is cAMP signalling.